HPLC法检查环戊丙酸雌二醇原料药的有关物质

目的:建立环戊丙酸雌二醇原料药中有关物质的检查方法。方法:采用高效液相色谱法。色谱柱为Kromasil C18,流动相为硝酸铵溶液-乙腈(23∶77,V/V),流速为1.5 ml/min,检测波长为280 nm,进样量为50μl。杂质Ⅰ按不加校正因子的主成分自身对照法计算,杂质Ⅱ采用杂质对照品外标法进行定量。结果:杂质Ⅰ与杂质Ⅱ检测质量浓度线性范围分别为0.3².0、0.52.0、0.5⁴.0μg/ml(r均为0.999 9),校正因子分别为0.970、0.368;杂质Ⅰ、杂质Ⅱ与环戊丙酸雌二醇检测限分别为6.3、1.5、2.3 ng,定量限分别为21.0、5.0、7.7 ng。3批样品中均检出杂质Ⅰ、杂质Ⅱ,但未检出雌二醇。结论:建立的方法准确、灵敏度高,可用于检查环戊丙酸雌二醇原料药中的有关物质。

Determination of Related Substances of Estradiol Cypionate Raw Material by HPLC

OBJECTIVE： To establish a method for the content determination of related substances in estradiol cypionate raw material. METHODS： HPLC method was adopted. The determination was performed on Kromasil C18 column with mobile phase consisted of ammonium nitrate-acetonitrile （23 ： 77, V/V） at the flow rate of 1.5 ml/min. The detection wavelength was set at 280 nm, and sample size was 50 μl. The impurity Ⅰ was determined by main component self contrast method without correction factor, and impurity Ⅱ was quantified by impurity control internal method. RESULTS： The linear range of impurity Ⅰ and impurity Ⅱ were 0.3-2.0 μg/ml （r=0.999 9） and 0.5-4.0 μg/ml （r=0.999 9）. The correction factors of impurity Ⅰ and impurity Ⅱ were 0.970 and 0.368. The detection limits of impurity Ⅰ , mpurity Ⅱ and estradiol cypionate were 6.3 ng, 1.5 ng and 2.3 ng, and the quantification limits were 21.0 ng, 5.0 ng and 7.7 ng, respectively. The impurity Ⅰ and impurity Ⅱ were detected in 3 batches of samples, and estradiol was not found. CONCLUSIONS： The method is accurate, sensitive and suitable for the determination of related substances in estradiol cypionate raw material.